Composition for stress relief containing fermented ripen noni and calamansi extract sugar as active ingredients

ABSTRACT

The present invention relates to a food composition for relieving stress, and specifically, to a food composition for relieving stress, which reduces secretion of the stress hormone cortisol and increases secretion of the happy hormone serotonin by containing fermented and aged noni and calamansi as active ingredients, has no side effects, and is safe for the human body. The composition of the present invention may be used as a health functional food or a pharmaceutical composition. The composition of the present invention has an excellent anti-stress effect.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of Korean Application Nos.10-2020-0185599, filed Dec. 29, 2020; and 10-2021-0166914, filed Nov.29, 2021; the entire contents of which are incorporated herein byreference.

BACKGROUND 1. Technical Field

The present invention relates to a composition containing fermented andaged noni and a calamansi extract as active ingredients, and moreparticularly, to a composition capable of relieving stress andinhibiting and alleviating stress-induced symptoms by containingfermented and aged noni and a calamansi extract as active ingredients.

2. Related Art

Stress, which is called the root of all diseases, exists in all areas oflife of humans and animals. Stress is a powerful stimulus, is induced byvarious factors ranging from trivial things experienced in daily life tomajor events that have a big impact. Stimulation by such stress inducesfatigue, malaise, depression, and the like, and causes internal andexternal negative reactions.

In general, if the stimulus on the body exceeds a certain degree, itcauses damage to the body, and in this case, the living body shows anon-specific general adaptive syndrome in response to a certain threatregardless of the type of stimulus, and this phenomenon is called stress(Selye, H.: The stress of life, Toronto, Longnans Green and Co., pp.1-50, 1958). Such stress causes various detrimental effects, such astension headache, migraine, hypertension, indigestion, fatigue, pain,menopausal disorders, hair loss, and rough skin. In addition, if stresscontinues chronically, it may cause various neuroses and gastric ulcers,reduce the physiological activity and immune activity of the body,impair mental activity, and make emotions unstable, resulting inabnormal behavior.

In particular, in modern society, stress excessively occurs, and thusstress has become a very important social problem.

It is known that the causes of stress and the changes in the bodythereby are diverse, but stress commonly causes hyperactivity of thehypothalamic-pituitary-adrenal (HPA) axis, which is the sympatheticnervous system, to induce hormone secretion, which causes hyperactivityof the adrenal gland and hypofunction of the spleen, resulting instress-induced symptoms. If stress is added, the weight of the spleen issignificantly reduced due to the decline in immune function,hyperactivity of the adrenal gland is caused by adrenocorticotropichormone and the weight thereof is increased, lipid reduction occurs, andthe secretion of cortisol is promoted. The amount of cortisol tends toincrease with age and stress, which also contributes to obesity. Inaddition, if stress is added, the adrenal glands secrete hormones,resulting in increases in cholesterol, glucose, and lactatedehydrogenase (LDH) levels in the blood.

Meanwhile, it is known that, in the case of gastric ulcers that aredeeply related to stress, reactive oxygen species are involved in theformation of ulcers. Biological defense systems for removing reactiveoxygen species (ROSs) are broadly classified into enzymatic defensesystems including enzymes such as superoxide dismutase (SOD), catalase,and glutathione peroxidase, and non-enzymatic defense systems that stopsor terminates the chain reaction of ROSs or free radicals. Organismsusing oxygen have SOD, which is an enzyme that removes superoxide, andthus the living body is protected from damage by superoxide. Catalasefunctions to protect the living body by removing H₂O₂ generated by theenzymatic reaction of SOD or the like in tissue. In addition, it hasbeen reported that catalase can not only remove free radicals ofreactive oxygen species generated during the metabolic process, but alsobe irreversibly inactivated by these reactive oxygen species, and alsoremoves free radicals generated by fatty acid oxidation. Glutathione(GSH), which is the most prevalent non-protein thiol in animal tissues,serve as a free radical scavenger and as a substrate for GSH-Px thatmetabolizes H₂O₂ and lipid peroxide, and plays an important role as anintracellular antioxidant (Cho S Y. et al., J. Korean Soc. Food Sci.Nutr., 32(3), pp. 458-463, 2003).

In recent years, various methods have been devised to relieve stress,and the most effective method is exercise or psychotherapy. In addition,there are methods for minimizing damage caused by stress by performingdrug therapy depending on the intensity of stress. In particular, drugsthat are used to relieve stress include psychotropic drugs such asbenzodiazepines, which are nerve stabilizers, and tranquilizer which aremental stabilizers.

However, when the above-described stabilizers are used for stressrelief, problems arise in that there is a risk of addiction and it isdifficult to exclude side effects, due to the characteristics of thesynthetic drugs. Excessive dependence on drugs has a negative side inthat it can cause other stress.

In recent years, various attempts have been made to develop foods forrecovering from fatigue or relieving stress using herbal medicinalmaterials.

As studies on compositions for recovering from fatigue using naturalherbal medicinal materials, Korean Patent Application Publication No.2000-0066777 discloses a functional beverage containing extracts fromCnidium officinale, licorice, Puerariae radix, Hovenia dulcis fruit,Alnus japonica, Citrus unshiu peel, Zizyphi fructus, Oenanthe javanica,and ginkgo leaf, and Korean Patent Application Publication No.10-2005-0108856 discloses a medicinal herbal drink for recovering fromfatigue containing Polygonatum sibiricum rhizome, Liriope platyphylla,Hovenia dulcis fruit, and Schisandra chinensis fruit.

As studies on foods for relieving stress, Korean Patent ApplicationPublication No. 10-2004-0006705 discloses a health supplement foodcontaining, as essential ingredients, deep water, organic germanium,Cyperus rotundus, Citrus unshiu peel, malt, Zizyphi spinosae semen, andjujube, and Korean Patent Application Publication No. 10-2004-0018888discloses a food for relieving stress containing, as active ingredients,red ginseng, Gastrodiae rhizome, and Rehmannia glutinosa juice. Inaddition, Korean Patent Application Publication No. 10-2005-0080455discloses Kami-sagunga-tang having an anti-stress effect, whichcontains, in addition to conventional Sagunga-tang, plum, Solomon'sseal, Puerariae radix, and cinnamon extracts.

It has been found that plants existing in nature contain large amountsof functional ingredients having bioregulatory functions, such asprevention of diseases and suppression of aging. Thus, studies onnatural food materials have been actively conducted.

Therefore, in recent years, studies have been actively conducted todevelop functional supplements using natural products with guaranteedsafety, such as plant extracts. Accordingly, the present inventors haveconducted intensive studies on fermented and aged noni and calamansi,and as a result, have found that intake of a complex of fermented andaged noni and calamansi is effective in relieving stress, therebycompleting the present invention.

PRIOR ART DOCUMENTS Patent Documents

(Patent Document 1) Korean Patent Application Publication No.2000-0066777

(Patent Document 2) Korean Patent Application Publication No.10-2005-0108856

SUMMARY

An object of the present invention is to solve the above-describedproblems and other problems related thereto.

The present inventors have conducted studies on a composition comprisingnatural ingredients capable of effectively relieving stress, and as aresult, have found that a composition containing a complex of fermentedand aged noni and a calamansi extract has a stress relief activity,thereby completing the present invention.

An object of the present invention is to provide a composition forrelieving stress containing, as active ingredients, fermented and agednoni or an extract thereof and a calamansi extract.

Another object of the present invention is to provide a method forproducing a composition containing, as active ingredients, fermented andaged noni or an extract thereof and a calamansi extract.

Still another object of the present invention is to provide a foodcomposition for relieving stress containing, as active ingredients,fermented and aged noni or an extract thereof and a calamansi extract.

Yet another object of the present invention is to provide apharmaceutical composition for relieving stress containing, as activeingredients, fermented and aged noni or an extract thereof and acalamansi extract.

Still yet another object of the present invention is to provide aquasi-drug composition for relieving stress containing, as activeingredients, fermented and aged noni or an extract thereof and acalamansi extract.

A further object of the present invention is to provide a feedcomposition for relieving stress containing, as active ingredients,fermented and aged noni or an extract thereof and a calamansi extract.

The objects to be achieved according to the technical idea of thepresent invention disclosed in the present specification are not limitedto the above-mentioned objects, and other objects not mentioned hereinwill be clearly understood by those skilled in the art from thefollowing description.

The present invention will be described in detail below. Meanwhile, eachdescription and embodiment disclosed in the present application may beapplied to each other description and embodiment. That is, allcombinations of various components disclosed in the present applicationfall within the scope of the present application. In addition, the scopeof the present application is not construed as being limited by thedetailed description described below.

To achieve the above-described objects, the present invention provides acomposition for relieving stress or inhibiting or treatingstress-induced diseases containing, as active ingredients, fermented andaged noni or an extract thereof and a calamansi extract.

Hereinafter, the present invention will be described in more detail.

As used herein, the term “fermented and aged noni” refers to noniobtained by fermenting and aging noni inoculated with lactic acidbacteria. Here, the part of noni that is used in the present inventionis not limited, but noni fruits may be used as an example.

In the present invention, “fermented noni” may be a fermented productobtained by fermenting noni fruits inoculated with any one or more ofthe following 7 types of lactic acid bacteria:

(1) Lactobacillus plantarum;

(2) Lactobacillus paracasei;

(3) Lactobacillus rhamnosus;

(4) Lactobacillus casei;

(5) Lactobacillus fermentum;

(6) Lactobacillus reuteri; and

(7) Lactococcus lactis subsp. lactis.

The “fermented noni” may be obtained by fermenting noni using lacticacid bacteria having excellent bioconversion ability so that theglycoside compounds scopolin and asperuloside contained in the noni maybe bioconverted to the non-glycoside compounds scopoletin,deacetylasperulosidic acid (DAA) and asperulosidic acid.

The content of bioconverted scopoletin in the “fermented noni” is 1 to200 μg/mL, the content of deacetylasperulosidic acid in the fermentednoni is 0.2 to 0.6 mg/mL, and the content of asperulosidic acid in thefermented noni is 0.096 to 1.41 mg/mL. The contents of scopoletin,deacetylasperulosidic acid and asperulosidic acid in the fermented noniare equal to those in fermented and aged noni within analysis errors.

In the present invention, “fermented and aged noni” may be fermented andaged noni itself or an extract obtained therefrom.

As an example, the fermented and aged noni of the present invention maybe obtained by fermenting and aging noni fruits inoculated with lacticacid bacteria, and the liquid obtained by squeezing the fermented andaged noni may be an extract.

Calamansi is a tropical fruit that is grown in Southeast Asia. The term“calamansi extract” means a fruit juice extracted from calamansi, andthe method of obtaining the fruit juice by extraction is well known inthe art, and thus detailed description thereof will be omitted. Inaddition, it is also possible to purchase and use a commerciallyavailable undiluted calamansi extract, rather than to prepare theextract separately. Calamansi has a 30 times higher vitamin C contentthan lemon, and contains large amounts of cryptoxanthin, hesperidin,nobiletin, cinehulin, and the like.

As used herein, the term “extract” commonly refers to a crude extract inthe art, but in a broad sense also refers to a fraction obtained byfractionating the extract. That is, the fermented and aged noni extractor the calamansi extract includes not only one obtained using anextraction solvent, but also one obtained by subjecting the obtained oneto a purification process. For example, the fermented and aged noniextract or calamansi extract of the present invention also includes afraction obtained by passing the extract through an ultrafiltrationmembrane with a predetermined molecular weight cut-off value, or afraction obtained by additionally performing various purificationmethods such as various chromatography methods (designed for separationaccording to size, charge, hydrophobicity, or hydrophilicity).

In the present specification, the expression “containing as activeingredients” means containing an amount sufficient to achieve theefficacy or activity of each of the fermented and aged noni or anextract thereof and the calamansi extract. In the present invention,since a composition containing an extract of each of the fermented andaged noni and calamansi, which are natural plant materials, and has noside effects on the human body even when the extracts are administeredin excessive amounts, the upper limit of the amount of each of thefermented and aged noni extract and calamansi extract contained in thecomposition of the present invention may be selected within anappropriate range by those skilled in the art.

Each of the fermented and aged noni extract and calamansi extractaccording to the present invention is obtained by extraction using anorganic solvent, and examples of an extraction solvent that may be usedin the present invention are as follows.

First, suitable examples of polar solvents include (i) water, (ii) analcohol having 1 to 6 carbon atoms (preferably, methanol, ethanol,propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol,2-butoxyethanol or ethylene glycol), (iii) acetic acid, (iv) DMFO(dimethyl formamide), and (v) DMSO (dimethyl sulfoxide).

Suitable examples of nonpolar solvents include acetone, acetonitrile,ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane,2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane,diisobutylene, 1-pentene, 1-chlorobutane, 1-chloropentane, o-xylene,diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane,chlorobenzene, benzene, diethyl ether, diethyl sulfide, chloroform,dichloromethane, 1,2-dichloroethane, aniline, diethylamine, ether,carbon tetrachloride, methylene chloride, petroleum ether, and THF.

Preferred examples of an extraction solvent that may be used in thepresent invention include (a) water, (b) an anhydrous or lower alcoholhaving 1 to 4 carbon atoms (e.g., methanol, ethanol, propanol, butanol,etc.), (c) a mixture of the lower alcohol and water, (d) acetone, (e)ethyl acetate, (f) chloroform, (g) butyl acetate, (h) 1,3-butyleneglycol, (i) hexane, and (j) diethyl ether. For easy extraction,extraction may be performed using water, ethanol, or a mixture of waterand ethanol.

In addition, each of the fermented and aged noni or an extract thereofand the calamansi extract, which are used in the present invention, maybe produced in a powder form by additional processes such asdistillation under reduced pressure and freeze-drying or spray-drying.

In the present invention, the fermented and aged noni or an extractthereof and the calamansi extract may be contained at a weight ratio of70:30 to 99:1, or 80:20 to 95:5, or 70:30 to 90:10.

According to one aspect of the present invention, the present inventionis directed to a food composition for relieving stress containing, asactive ingredients, fermented and aged noni or an extract thereof and acalamansi extract.

The food composition may include, for example, a health functional foodcomposition. Examples of the health functional food composition includevarious foods, beverages, gum, tea, vitamin complexes, healthsupplements, etc. The health functional food composition may be used inthe form of powders, granules, pills, tablets, capsules, candies, syrupsor beverages. The food composition of each formulation may contain, inaddition to the active ingredient, ingredients commonly used in the art,which may be appropriately selected by those skilled in the art withoutdifficulty depending on the formulation or intended use of thecomposition. When the food composition is applied simultaneously withother raw materials, a synergistic effect may occur.

The composition may contain other ingredients, which may impart asynergistic effect to the main effect, within a range that does notimpair the main effect of the present invention. For example, thecomposition may further contain additives such as fragrance, a colorant,a bactericide, an antioxidant, a preservative, a humectant, a thickener,an inorganic salt, an emulsifier, and a synthetic polymer, in order toimprove physical properties thereof.

When the health functional food composition of the present invention isused as a food additive, the fermented and aged noni or an extractthereof and the calamansi extract may be added as they are or usedtogether with other foods or food ingredients, and may be appropriatelyused according to a conventional method. The content of each of theactive ingredients may be suitably determined according to the intendeduse (prevention, health improvement or therapeutic treatment). The foodis not particularly limited in the kind thereof. Examples of food towhich the fermented and aged noni or an extract thereof and thecalamansi extract may be added include meat, sausage, bread, chocolate,candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairyproducts including ice cream, various soups, beverages, teas, drinks,alcoholic beverages, vitamin complexes, etc., and include all healthfunctional foods in the ordinary sense.

When the composition of the present invention is used as a health drink,it may contain various flavoring agents or natural carbohydrates asadditional ingredients, like a conventional drink. Examples of theabove-described natural carbohydrates include monosaccharides such asglucose and fructose, disaccharides such as maltose and sucrose,polysaccharides such as dextrin and cyclodextrin, and sugar alcoholssuch as xylitol, sorbitol and erythritol. As the flavoring agents, theremay be used natural flavoring agents such as thaumatin or steviaextract, or synthetic flavoring agents such as saccharin or aspartame.

The present invention provides a method for producing a compositioncontaining fermented and aged noni and calamansi, the method comprisingsteps of: obtaining fermented and aged noni by fermenting and aging noniusing lactic acid bacteria; and adding and mixing calamansi with thefermented and aged noni.

According to one aspect of the present invention, the present inventionprovides a method for producing a composition containing fermented andaged noni or an extract thereof and a calamansi extract, the methodcomprising steps of: (a) obtaining fermented and aged noni by fermentingand aging noni using lactic acid bacteria; and (b) adding calamansi tothe fermented and aged noni, followed by aging.

According to one aspect of the present invention, the present inventionprovides a method for producing a composition for relieving stress, themethod comprising steps of: (a) producing bioconverted fermented andaged noni by fermenting and aging noni fruits inoculated with lacticacid bacteria; (b) squeezing the fermented and aged noni; and (c) addingand mixing calamansi with the fermented and aged noni.

The fermentation and aging may be performed at 35 to 40° C. for 48 to168 hours.

The aging may be performed by pouring the fermentation broth, producedin the fermentation process, over the fermentation product at regulartime intervals.

The bioconverted fermented and aged noni may be a fermented productobtained by fermenting noni fruits inoculated with any one or more ofthe above-described 7 types of lactic acid bacteria.

According to one aspect of the present invention, the present inventionis directed to a pharmaceutical composition for relieving stress orinhibiting or treating stress-induced diseases containing, as activeingredients, fermented and aged noni or an extract thereof and acalamansi extract.

In an example of the present invention, it was confirmed that, when thepharmaceutical composition of the present invention was used, thesecretion of the stress hormone cortisol was suppressed. Thus, it can beseen that the pharmaceutical composition of the present invention mayalso inhibit stress-induced behavioral changes, and thus exhibit theeffect of inhibiting or treating stress-induced diseases such asdepression, anxiety disorder, fatigue syndrome, sleep disorder, panicdisorder, memory loss, lethargy, insomnia, Parkinson's disease, andAlzheimer's disease.

Therefore, the present invention is directed to a pharmaceuticalcomposition for inhibiting or treating stress-induced diseasescontaining, as active ingredients, fermented and aged noni or an extractthereof and a calamansi extract.

The pharmaceutical composition according to the present invention maycontain the active ingredients alone or may be formulated in a suitableform together with a pharmaceutically acceptable carrier, and mayfurther contain an excipient or a diluent. As used herein, the term“pharmaceutically acceptable” refers to a non-toxic composition which isphysiologically acceptable and, when administered to the human beings,does not cause allergic reactions such as gastrointestinal disorders anddizziness, or similar reactions.

The pharmaceutical composition may further contain a pharmaceuticallyacceptable carrier such as a carrier for oral administration or acarrier for parenteral administration. Examples of the carrier for oraladministration include lactose, starch, cellulose derivatives, magnesiumstearate, stearic acid, and the like. In addition, various drug deliverymaterials that are used for oral administration of peptide formulationsmay be included. In addition, examples of the carrier for parenteraladministration include water, suitable oil, saline, aqueous glucose andglycol, and the like, and further include a stabilizer and apreservative. Suitable stabilizers include antioxidants such as sodiumhydrogen sulfite, sodium sulfite or ascorbic acid. Suitablepreservatives include benzalkonium chloride, methyl- or propyl-parabenand chlorobutanol. The pharmaceutical composition of the presentinvention may further contain a lubricant, a wetting agent, a sweeteningagent, a flavoring agent, an emulsifying agent, a suspending agent, andthe like, in addition to the above-described ingredients. Otherpharmaceutically acceptable carriers and agents may refer to thosedescribed in Remington's Pharmaceutical Sciences, 19th ed., MackPublishing Company, Easton, Pa., 1995.

The composition of the present invention may be administered to mammalsincluding humans by any method. For example, the composition may beadministered orally or parenterally. Parenteral administration methodsinclude, but are not limited to, intravenous, intramuscular,intraarterial, intramedullary, intrathecal, intracardiac, transdermal,subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingualor rectal administration.

The pharmaceutical composition of the present invention may beformulated in an oral dosage form or a parenteral dosage form dependingon the administration route as described above.

For oral dosage forms, the composition of the present invention may beformulated into a powder, granule, tablet, pill, dragee, capsule,liquid, gel, syrup, slurry, suspension, etc. using methods known in theart. For example, the oral dosage form may be obtained as a tablet ordragee by mixing the active ingredient with a solid excipient,pulverizing the mixture, adding a suitable adjuvant thereto, andprocessing the mixture into a granule mixture. Examples of suitableexcipients include sugars such as lactose, dextrose, sucrose, sorbitol,mannitol, xylitol, erythritol and maltitol, starches such as cornstarch, wheat starch, rice starch and potato starch, celluloses such ascellulose, methyl cellulose, sodium carboxymethyl cellulose andhydroxypropylmethyl cellulose, and fillers such as gelatin andpolyvinylpyrrolidone. In addition, if necessary, cross-linkedpolyvinylpyrrolidone, agar, alginic acid or sodium alginate may be addedas a disintegrant. Furthermore, the pharmaceutical composition of thepresent invention may further contain an anti-aggregating agent, alubricant, a wetting agent, a flavoring agent, an emulsifying agent, anda preservative.

For parenteral dosage forms, the pharmaceutical composition may beformulated in the form of an injection, cream, lotion, externalointment, oil, humectant, gel, aerosol or nasal inhalant by methodsknown in the art. These formulations are described in Remington'sPharmaceutical Science, 19^(th) ed., Mack Publishing Company, Easton,Pa., 1995, which is a prescription commonly known in all pharmaceuticalchemistries.

The total effective amount of the composition of the present inventionmay be administered to a patient in a single dose, and may beadministered in multiple doses by a fractionated treatment protocol overa prolonged period of time. The pharmaceutical composition of thepresent invention may have varying contents of the active ingredientdepending on the severity of the disease. Preferably, the preferredtotal dose of the pharmaceutical composition of the present inventionmay be about 0.01 μg to 10,000 mg, preferably 0.1 μg to 500 mg, mostpreferably 100 mg to 500 mg per kg of patient body weight per day.However, the effective dose of the pharmaceutical composition to beadministered to a patient is determined in consideration of variousfactors, including the formulation method, the route of administrationand the number of treatments, as well as the patient's age, weight,health status and sex, the severity of the disease, diet, and excretionrate, and thus those of ordinary skill in the art will be able todetermine an appropriate effective dosage of the composition of thepresent invention in consideration of these factors. The pharmaceuticalcomposition according to the present invention is not particularlylimited in its formulation, administration route and administrationmethod, as long as it exhibits the effect of the present invention.

According to one aspect of the present invention, the present inventionis directed to a quasi-drug composition for relieving stress orinhibiting or treating stress-induced diseases containing, as activeingredients, fermented and aged noni or an extract thereof and acalamansi extract.

As used herein, the term “quasi-drug” refers to a product which, amongthe products being used for the purpose of treatment, alleviation,handling, or prevention of human or animal diseases, excludes thosewhich are not a tool, a machine, or a device, and a product which, amongthe products being used for the purpose of rendering a pharmacologicaleffect on the human or animal structures and functions, excludes thosewhich are not a tool, a machine, or a device. In one embodiment, thequasi-drug may include an agent for internal administration, but is notlimited thereto, and the formulation method, dose, use method,components, etc. of the quasi-drug may be appropriately selected usingconventional techniques known in the art.

The quasi-drug composition of the present invention may further contain,in addition to the above ingredient, a pharmaceutically acceptablecarrier, excipient or diluent, if necessary. The pharmaceuticallyacceptable carrier, excipient or diluent is not limited as long as itdoes not impair the effects of the present invention, and examplesthereof include a filler, an extender, a binder, a wetting agent, adisintegrant, a surfactant, a lubricant, a sweetener, a fragrance, and apreservative.

According to one aspect of the present invention, the present inventionis directed to a feed composition for relieving stress or inhibiting ortreating stress-induced diseases containing, as active ingredients,fermented and aged noni or an extract thereof and a calamansi extract.

In the present invention, the feed composition may be formulated in aconventional feed form, and may also contain known feed ingredients. Inaddition, the feed composition may be used as a feed additive, which isadded in the form of an additive to the feed used. The feed additive ofthe present invention corresponds to a supplementary feed under the FeedManagement Act, and may further contain minerals such as sodiumbicarbonate (soda), bentonite, magnesium oxide, and composite minerals,trace minerals such as zinc, copper, cobalt, and selenium, vitamins suchas carotene, vitamin E, vitamins A, D, E, nicotinic acid, and vitamin Bcomplexes, protective amino acids such as methionine and lysine,protective fatty acids such as fatty acid calcium salts, live bacteriasuch as probiotics (lactic acid bacteria), yeast cultures, and fungalfermentation products, yeast agents, etc.

The feed or feed additive of the present invention may be applied todiets for a number of animals, including mammals, poultry and fish.

The fermented and aged noni or extract thereof and calamansi extractcontained in the composition for relieving stress according to thepresent invention may be used as a natural pharmaceutical composition orfood composition, and has the effects of relieving stress and inhibitingor efficiently alleviating stress-induced symptoms (diseases) withoutside effects.

In addition, the fermented and aged noni contains bioconvertedcomponents (scopoletin, deacetylasperulosidic acid and asperulosidicacid), which are obtained in the process of fermenting noni fruits andare capable of improving digestion and absorption, and thus it promotesdigestion, absorption, and bowel movement in adults and the elderly whohave relatively low digestibility. In addition, since the fermented andaged noni is a natural raw material, it has no toxicity or side effectsand may be safe to consume.

In addition, the process of producing the fermented and aged noni issimple, and thus may greatly reduce the production cost, thereby greatlyreducing the cost burden of consumers.

In addition, the composition containing the complex of the fermented andaged noni and the calamansi extract has improved sensory characteristicscompared to conventional noni, and thus is a food composition thatconsumers can enjoy without burden.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the results of measuring the immobility timesof experimental animals in a forced swimming test after intake of eachof a control, a fermented and aged noni extract, a calamansi extract,and complexes of the fermented and aged noni extract and the calamansiextract in an experimental example of the present invention.

FIG. 2 is a graph showing the results of measuring the immobility timesof experimental animals in a tail suspension test after intake of eachof a control, a fermented and aged noni extract, a calamansi extract,and complexes of the fermented and aged noni extract and the calamansiextract in an experimental example of the present invention.

FIG. 3 is a graph showing the results of testing changes in the serumIgA levels in immobilization stress-induced mice after intake of each ofa control, a fermented and aged noni extract, a calamansi extract, andcomplexes of the fermented and aged noni extract and the calamansiextract in an experimental example of the present invention.

FIG. 4 is a graph showing the results of testing changes in the bodyweights of immobilization stress-induced mice after intake of each of acontrol, a fermented and aged noni extract, a calamansi extract, andcomplexes of the fermented and aged noni extract and the calamansiextract in an experimental example of the present invention.

FIG. 5 is a graph showing the results of testing changes in the serumserotonin levels in immobilization stress-induced mice after intake ofeach of a control, a fermented and aged noni extract, a calamansiextract, and complexes of the fermented and aged noni extract and thecalamansi extract in an experimental example of the present invention.

FIG. 6 is a graph showing the results of testing changes in the serumcortisol levels in immobilization stress-induced mice after intake ofeach of a control, a fermented and aged noni extract, a calamansiextract, and complexes of the fermented and aged noni extract and thecalamansi extract in an experimental example of the present invention.

DETAILED DESCRIPTION

Hereinafter, the present invention will be described in more detail withreference to the following examples. However, these examples servemerely to illustrate the present invention, and the scope of the presentinvention is not limited to these examples.

EXAMPLE 1 Production of Compositions for Relieving Stress

Compositions according to the present invention were produced throughthe following process.

1.1: Production of Fermented and Aged Noni

690 kg of noni fruits were inoculated with complex lactic acid bacteria(a complex of 7 types of lactic acid bacteria, AON1805, Lactomason Co.,Ltd.) and fermented at 37° C. for 45 days or more. Fermentation andaging were performed while 10 L of the fermentation broth collectedevery 7 days from the start date of fermentation was poured over thenoni fruits. The composition of the complex lactic acid bacteria (acomplex of 7 types of lactic acid bacteria, AON1805, Lactomason Co.,Ltd.) used is shown in Table 1 below.

After completion of the aging, the fermented and aged noni was squeezedand the solid was removed, thus obtaining a fermented and aged noniextract.

TABLE 1 Raw material name or ingredient name and mixing ratio No. Rawmaterial name or ingredient name Mixing ratio (%) 1 Lactobacillusplantarum 70% 2 Lactobacillus rhamnosus  5% 3 Lactobacillus casei  5% 4Lactobacillus ferrnenturn  5% 5 Lactobacillus paracasei  5% 6Lactobacillus reuteri  5% 7 Lactococcus lactis subsp. lactis  5%

1.2: Production of Calamansi Extract

As calamansi, calamansi sold in the market was purchased and used inthis Example. As frozen or dried calamansi, frozen or dried calamansisold in the market was purchased and used.

The frozen calamansi was pulverized and squeezed, thus producing anextract. Meanwhile, 100 g of the dried calamansi was extracted twice at50° C. for 8 hours after a 10-fold amount (weight) of water was addedthereto, and then the extract was filtered through filter paper. Thefiltrate was dried using a rotary vacuum evaporator (EYELA A-1000S,Tokyo Rikakikai Co., Tokyo, Japan) on a water bath at 37 to 40° C. toremove water, thus producing an extract, which was used as a sample.

The yields of the calamansi extract samples obtained using differentextraction solvents are shown in Table 2 below.

TABLE 2 Sample Extract Yield (%) Dried calamansi extract (DCE) Waterextract 13.2 Frozen calamansi extract (FCE) Squeezed extract 15.3

1.3: Production of Complexes Using Fermented and Aged Noni Extract andCalamansi Extract

The fermented and aged noni extract produced in Example 1.1 and thecalamansi extract produced in Example 1.2 were mixed together at variousratios and then aged for 15 days to obtain complexes.

[Complex Production]

Complex 1: 90 wt % fermented and aged noni extract: 10 wt % driedcalamansi extract

Complex 2: 80 wt % fermented and aged noni extract: 20 wt % driedcalamansi extract

Complex 3: 70 wt % fermented and aged noni extract: 30 wt % driedcalamansi extract

Complex 4: 90 wt % fermented and aged noni extract: 10 wt % frozencalamansi extract

Complex 5: 80 wt % fermented and aged noni extract: 20 wt % frozencalamansi extract

Complex 5: 70 wt % fermented and aged noni extract: 30 wt % frozencalamansi extract

EXPERIMENTAL EXAMPLE 1 Evaluation of the Effect of Composition ofPresent Invention on Stress Relief

An evaluation was performed of the stress relief effects of thefermented and aged noni extract produced in Example 1.1, the calamansiextract produced in Example 1.2, and the complexes produced in Example1.3.

(1) Experimental Mice

4-week-old ICR mice (Orient Bio, Korea) were adapted for 1 week at atemperature of 18° C. to 23° C. and a humidity of 60% with a 12-hrlight/12-hr dark cycle. Then, the experimental animals were divided intoexperimental groups (8 mice per group) as follows. The mice were fedsolid feed, and allowed to access feed and water ad libitum, except forthe process of inducing stress.

Experimental group 1 (normal control group, hereinafter referred to asNormal); experimental group 2 (negative control group; hereinafterreferred to as Control); experimental group 3 (a group to which 100mg/kg of the fermented and aged noni was administered; hereinafterreferred to as FN); experimental group 4 (a group to which 100 mg/kg ofthe dried calamansi extract was administered; hereinafter referred to asDCE); experimental group 5 (a group to which 100 mg/kg of the frozencalamansi extract was administered; hereinafter referred to as FCE);experimental group 6 (a group to which 100 mg/kg of complex 1 wasadministered; hereinafter referred to as C 1), experimental group 7 (agroup to which 100 mg/kg of complex 2 was administered; hereinafterreferred to as C 2); experimental group 8 (a group to which 100 mg/kg ofcomplex 3 was administered; hereinafter referred to as C 3);experimental group 9 (a group to which 100 mg/kg of complex 4 wasadministered; hereinafter referred to as C 4); experimental group 10 agroup to which 100 mg/kg of complex 5 was administered; hereinafterreferred to as C 5); and experimental group 11 (a group to which 100mg/kg of complex 6 was administered; hereinafter referred to as C 6).Each test substance was administered once a day for a total of 4 weeks.

(2) Stress Induction

Stress was induced every day for 4 weeks from 7 days afteradministration of physiological saline to the negative control group andadministration of the extract alone or each complex to experimentalgroup 1, experimental group 2, experimental group 3, experimental group4, experimental group 5 and experimental group 6. Stress induction wasperformed using a modification of the method of Willner et al. (Wilneret al., Reduction of sucrose preference by chronic unpredictable mildstress, 1987). Specifically, stress was induced by creating variousunexpected psychological stressful situations, such as fasting, dietaryrestriction after fasting (feeding in small amounts), water supply cutoff, provision of empty water bottles after water supply cut off,slanted cages, breeding of multiple experimental animals in one cage,flashing light, cold room, and continuous lighting.

(3) Forced Swimming Test

The experimental animals were subjected to a forced swimming test. Aftera cylindrical transparent water bath (14 cm diameter and 20 cm height)was filled with water at 23 to 25° C. so that the water depth was 15 cm,each experimental animal was put into the water bath. Each experimentalanimal was forced to swim in the cylindrical water bath for 6 minutes. 2minutes after the start was considered an adaptation time, and during 4minutes before the end, the total time during which each experimentalanimal was immobile in the cylindrical water bath (hereinafter referredto as “immobility time”) was measured and recorded.

Specifically, the term “immobility time” refers to a state (immobility)in which there is no movement except for small movements such as raisingthe head above the water surface. Immobility is one of the indicators ofdepression, and can be commonly seen in stress-induced animals.

As shown in FIG. 1, it was confirmed that the immobility time of thenegative control group (Control) was longer than that of the normalcontrol group (Normal). On the other hand, it can be seen that theimmobility time of experimental group 3 was slightly longer than that ofthe normal control group, but was shorter than (about 80% of) that ofthe negative control group. However, it was confirmed that theimmobility times of experimental groups 4 and 5 showed a significantdifference from that of the negative control group, and the immobilitytimes of experimental groups 6, 7, 8, 9, 10 and 11, to which thecomplexes of the fermented and aged noni extract and the calamansiextract were administered, significantly decreased to about half of thatof the negative control group. Therefore, it can be seen that thecomplex is more effective in reducing immobility time than the fermentedand aged noni extract alone or the calamansi extract alone.

(4) Tail Suspension Test

The experimental animals were subjected twice to a tail suspension test.Each experimental animal was hung upside down for 6 minutes by holdingthe 1 cm tip of the tail at a height of 15 cm from the bottom, and themovement of the experimental animal was observed. 2 minutes after thestart was considered an adaptation time, and during 4 minutes before theend, the time during which each experimental animal was immobile wasmeasured and recorded.

As shown in FIG. 2, it was confirmed that the immobility time of thenegative control group (Control) in the state in which the animal washung upside down was longer than that of the normal control group(Normal). This means that the immobility time was increased due tomental stress, suggesting that the frustration of the animals of thenegative control group increased. The immobility time of experimentalgroup 3 (FN) was short than (about 82% of) than that of the negativecontrol group, suggesting that experimental group 3 tried to move thebody relatively, but the immobility time of experimental group 4 wasabout 92% of that of the negative control group, which is a significantdifference.

In addition, it was confirmed that the immobility times of the groups,to which each of complexes 1, 2, 3, 4, 5 and 6 produced by mixing thefermented and aged noni extract (FN) and the calamansi extract (DCE orFCE) together at a predetermined ratio was administered, were allsignificantly shorter than that of the negative control group.Therefore, it can be seen that the complex obtained by mixing thefermented and aged noni extract and the calamansi extract together at apredetermined ratio is more effective in alleviating stress inducedlethargy and frustration.

EXPERIMENTAL EXAMPLE 2 Test for Changes in Serum IgA Levels inImmobilization Stress-Induced Mice After Intake of Composition

To immobilization stress-induced 8 week-old C57BL/6 mice in which stresswas induced by immobilizing the mice in narrow frames for 2 hours everyday for a total of 21 days, each of the fermented and aged noni extract(FN), the calamansi extract (CE), and complexes 1, 2, 3, 4, 5 and 6,produced by mixing the fermented and aged noni extract (FN) and thecalamansi extract (DCE or FCE) together at a predetermined ratio, wasorally administered every day at a dose of 100 mg/kg.

As shown in FIG. 3, as a result of measuring the serum IgA levels, itwas confirmed that the serum IgA levels in the groups, to which each ofthe fermented and aged noni extract (FN), complex 1 and complex 4 wasadministered, statistically significantly increased compared to that inthe control group (Control).

In particular, the group to which complex 4 was administered showed thehighest increase in the serum IgA level.

EXPERIMENTAL EXAMPLE 3 Test for Changes in Body Weights ofImmobilization Stress-Induced Mice After Intake of Composition

To immobilization stress-induced 8 week-old C57BL/6 mice in which stresswas induced by immobilizing the mice in narrow frames for 2 hours everyday for a total of 21 days, each of the fermented and aged noni extract(FN), the calamansi extract (CE), and complexes 1, 2, 3, 4, 5 and 6,produced by mixing the fermented and aged noni extract (FN) and thecalamansi extract (DCE or FCE) together at a predetermined ratio, wasorally administered every day at a dose of 100 mg/kg.

As shown in FIG. 4, as a result of measuring the changes in body weightof the mice, it was confirmed that the immobilization stress-inducedcontrol group (Control) showed a weight loss compared to the normalgroup (Normal), and the weight losses of the groups, to which each ofthe fermented and aged noni extract (FN), complex 1 and complex 4 wasadministered, were smaller than that of the immobilizationstress-induced control group (Control).

In particular, the groups to which each of complexes 1 and 4 wasadministered showed the lowest weight loss.

EXPERIMENTAL EXAMPLE 4 Test for Changes in Serum Cortisol and SerotoninLevels in Immobilization Stress-Induced Mice After Intake of Composition

To immobilization stress-induced 8 week-old C57BL/6 mice in which stresswas induced by immobilizing the mice in narrow frames for 2 hours everyday for a total of 21 days, each of the fermented and aged noni extract(FN), the calamansi extract (CE), and complexes 1, 2, 3, 4, 5 and 6produced by mixing the fermented and aged noni extract (FN) and thecalamansi extract (DCE or FCE) together at a predetermined ratio wasorally administered every day at a dose of 100 mg/kg.

As a result of measuring the serum level of serotonin which is a hormonethat makes the body feel happy, as shown in FIG. 5, it was confirmedthat the groups, to which each of the fermented and aged noni extract(FN), complex 1 and complex 4 was administered at a dose of 100 mg/kg,showed a statistically significant increase in the serotonin levelcompared to the immobilization stress-induced group (Control) (p<0.05).

As a result of measuring the serum level of cortisol that is a stresshormone, as shown in FIG. 6, it was confirmed that the groups, to whicheach of the fermented and aged noni extract (FN), complex 1 and complex4 was administered, showed a statistically significant decrease in thecortisol level compared to the immobilization stress-induced group(Control) (p<0.05).

In particular, the groups to which each of complexes 1 and 4 wasadministered showed the highest increase in the serotonin level and thehighest decrease in the cortisol level.

While the present invention has been described with reference to theparticular illustrative embodiments, those of ordinary skill in the artto which the present invention pertains will appreciate that the presentinvention may be embodied in other specific forms without departing fromthe technical spirit or essential characteristics of the presentdisclosure. Therefore, the embodiments described above are considered tobe illustrative in all respects and not restrictive. The scope of thepresent invention should be defined by the claims rather than theforegoing description, and it should be understood that allmodifications or variations derived from the meanings and scope of thepresent invention and equivalents thereof are included in the scope ofthe appended claims.

What is claimed is:
 1. A food, pharmaceutical, quasi-drug, or feed composition for relieving stress or inhibiting or treating stress-induced diseases containing, as active ingredients, fermented and aged noni or an extract thereof and a calamansi extract.
 2. The food, pharmaceutical, quasi-drug, or feed composition of claim 1, wherein the fermented and aged noni or extract thereof and the calamansi extract are contained at a weight of 70:30 to 99:1.
 3. The food, pharmaceutical, quasi-drug, or feed composition of claim 1, wherein the fermented and aged noni or extract thereof is obtained by fermenting and aging noni using lactic acid bacteria.
 4. The food, pharmaceutical, quasi-drug, or feed composition of claim 1, which is a health functional food.
 5. The food, pharmaceutical, quasi-drug, or feed composition of claim 1, which has any one formulation selected from among powders, granules, pills, tablets, capsules, candy, syrups and beverages.
 6. A pharmaceutical composition according to claim
 1. 7. A quasi-drug composition according to claim
 1. 8. A feed composition according to claim
 1. 9. A method for producing a composition containing fermented and aged noni or an extract thereof and a calamansi extract, the method comprising steps of: (a) obtaining fermented and aged noni by fermenting and aging noni using lactic acid bacteria; and (b) adding calamansi to the fermented and aged noni, followed by aging.
 10. The method of claim 9, further comprising, after step (a), a step of obtaining a fermented and aged noni extract by squeezing the fermented and aged noni obtained in step (a).
 11. A composition obtained according to the method of claim
 9. 12. The composition of claim 11, which has a stress relief ability. 